The heading-date gene Ghd7 inhibits seed germination by modulating the balance between abscisic acid and gibberellins

Seed dormancy of cultivated rice was largely weakened during the progress of domestication.Correct timing and uniformity of seed germination are important for rapid seedling establishment and highyield production.In the present study,we found that the heading-date gene Ghd7 acted as a negative regulator of germination.A mutant of ghd7 showed low sensitivity to exogenous ABA treatment during seed germination.Further investigation revealed reduced accumulation of ABA in mature ghd7 seeds as a consequence of dampened expression of OsNCED genes.Moreover,elevated GA_(3) level was detected in seeds of ghd7 mutant during imbibition course,which was attributed to the induction of genes responsible for the synthesis pathways of bioactive GAs.Thus,Ghd7 inhibits seed germination by increasing the ABA/GA_(3) ratio.Besides revealing pleiotropic effects of Ghd7,our results indicate its role in linking seed germination to growth-phase transition in rice,which would enrich the theoretical basis for future breeding practices.

BEAR1, a bHLH transcription factor, controls seedling growth by regulating gibberellins biosynthesis in rice

The genetic pathways of rice seedling growth have a major impact on seedling emergence from soil and development.In this study,we identified a new bHLH transcription factor,BEAR1,from rice RNAi mutant library.Both the BEAR1-RNAi and bear1 CRISPR mutants had accelerated seedling growth.Histological section of leaves showed accelerated development of lacuna and vascular bundles in bear1 mutant.GUS staining revealed that BEAR1 was highly expressed in coleoptiles and leaves at seedling stage.Expression analysis of gibberellin(GA)biosynthesis and metabolic genes and content determination of active GAs indicated that the expression of GA biosynthesis genes,especially OsKS4 and OsCPS2,were upregulated and the GAs content were significantly increased in bear1,which correlated with the seedling phenotype of bear1 mutant.Molecular and biochemical assays revealed that BEAR1 directly binds to the promoter of OsKS4,thereby repressing its expression.Haplotypes analysis showed clear differentiation in indica and japonica rice varieties,and a strong correlation with seedling height.These findings provide novel insights into the regulation of seedling growth in rice.

Seed Germination in Tomato: A Focus on Interaction between Phytochromes and Gibberellins or Abscisic Acid

Separately, it is well-documented that phytochromes (phys), gibberellin (GA) and abscisic acid (ABA) strongly control the seed germination in tomato. However, we hipothesized that phys interact with GA or ABA during this response. Thus, we make an analysis of seed germination of ABA deficient (sit), GA constitutive response (pro), phytochrome deficient (au) mutants as well as, specially, au sit and au pro double mutants of tomato?incubated in the dark or light conditions during 120 h [12 h intervals (i)]. Compared to au, which severely?reduced percentage germination (Gi%) and pro, which did not alter Gi%, au pro showed in the light enhanced Gi% and germination speed index (GSI) besides the reduced average germination time (AGT). Moreover, in the dark, germination of au pro was similar to pro. These results indicate that the mechanisms by which GA modulate germination in tomato are light dependent through the phy signaling, whereas intermediary values of Gi%, GSI and AGT in dark and light of au sit compared to au and sit single mutants indicate an additive effect of the au and sit mutations, suggesting that ABA and phy may act through the parallel signaling pathway.

Gibberellins,brassinolide,and ethylene signaling were involved in flower differentiation and development in Nelumbo nucifera

Hormones play important roles in vegetative and reproductive processes;however,the regulatory roles of hormones in Nelumbo nucifera(Lotus)growth and development are unclear.In this study,nine types of endogenous hormones,including gibberellins(GA_1,GA_3,and GA_4),indole-3-acetic acid(IAA),brassinolide(BR),ethylene(ETH),jasmonic acid(JA),abscisic acid(ABA),and zeatin(ZT)were detected in dormant shoot tips,vegetative shoot tips,developing leaf buds,and developing flower buds of lotus.The results indicated that GA,ETH,and BR signaling can promote vegetative and reproductive development of lotus.GA signaling regulates plant height and stimulates flower bud differentiation.GA levels were the highest in the flower buds;exogenous GA3+4 increased plant height by approximately 90%,increased flower quantity by nearly 40%,and advanced flowering by 4 d.Suppressing GA biosynthesis using paclobutrazol decreased plant height and flower quantity by 38%and 87.1%,respectively,and delayed flowering by 15.6 d.ETH signaling has positive regulatory effects on vegetative growth and flower development.The ETH concentration in the developing leaf buds was at least 50%higher than that in other samples.Ethephon spraying led to remarkable increases in plant height and leaf thickness and extended the flowering duration.BR signaling acts as a growth promoter during vegetative and reproductive development in lotus.The highest BR levels were detected in the vegetative shoot tips.External application of28-epihomobrassinolide resulted in growth-promoting phenotypes including longer scapes,thicker leaves,and prolonged flowering.

Response of Two Varieties Eggplant (Solanum melongena L.) Growth and Production to Many Kinds of Gibberellins

The aim of this study was to know the influence of plant growth regulator gibberellins on plant growth and production of Kopek and Gelatik purple eggplant.This study has been done in Educational and Research Garden in Stiper Agriculture University at Maguwoharjo,Depok,Sleman,Yogyakarta in September 2016 to February 2017.The experimental design used was CRD(Completely Randomized Design)with two factors.The first factor was different kinds of gibberellins consisting of G0(no gibberellin/control),G1(young bamboo gibberellin)and G2(corn gibberellin)and G3(inorganic gibberellin).The second factor was the kind of 2 varieties consisting of purple eggplants.V1 variety was Kopek purple eggplant and V2 variety was Gelatik purple eggplant.The study used six replications.Data were analyzed by Anova followed by DMRT(Duncan’s Multiple Range Test)at 5%significant level.The result showed significantly interaction between kinds of gibberellins with eggplant variety on plant height.The Kopek purple with inorganic gibberellin significantly was the best.Kopek purple eggplant was much more in growth and production than Gelatik purple eggplant.

Light Regulation of Gibberellins Metabolism in Seedling Development

Light affects many aspects of plant development, including seed germination, stem elongation, and floral initiation. How photoreceptors control photomorphogenic processes is not yet fully understood. Because phytohormones are chemical regulators of plant development, it may not be surprising that light affects, directly or indirectly, cellular levels and signaling processes of various phytohormones, such as auxin, gibberellins （GA）, cytokinin, ethylene, abscisic acid （ABA）, and brassinosteroids （BR）. Among those phytohormones, light regulation of GA metabolism has probably attracted more attention among photoblologists and it is arguably the most extensively studied plant hormone at present with respect to its role in photomorphogenesis. It has become Increasingly clear that phytochromes and cryptochromes are the major photoreceptors mediating light regulation of GA homeostasis. This short article attempts to examine some recent developments in our understanding of how light and photoreceptors regulate GA blosynthesis and catabolism during seedling development. It is not our intention to carry out a comprehensive review of the field, and readers are referred to recent review articles for a more complete view of this area of study （Kamiya and Garcia-Martinez 1999; Hedden and Phillips 2000; Garcla-Martinez and GI12001; Olszewski et al. 2002; Halliday and Fankhauser 2003; Sun and Gubler 2004）.

Phytohormones Jasmonic Acid,Salicylic Acid,Gibberellins,and Abscisic Acid are Key Mediators of Plant Secondary Metabolites

Until recently,many studies on the role of phytohormones in plant secondary metabolism focused on jasmonic acid(JA),salicylic acid(SA),gibberellins(GA),and abscisic acid(ABA).It is now clear that phytohormone?induced regulation of signaling occurs via regulation of the biosynthetic pathway genes at the transcriptional level or through posttranslational regulation,or an increase in secondary metabolite deposition(e.g.,trichomes).Here,we summarize recent advances,updating the current reports on the molecular machinery of phytohormones JA,SA,GA,and ABA involved in plant secondary metabolites.This review emphasizes the differences and similarities among the four phytohormones in regulating various secondary metabolic biosynthetic pathways and also provides suggestions for further research.

Concurrent Deficiency of Gibberellins and Abscisic Acid Causes Plant Male Sterility

Plant fertility influences crop yield and quality, and thus is an important economic trait for agricultural production. There- fore, it is essential to understand the precise molecular mechanisms that control plant fertility. In the past decades, many studies demonstrated that the phytohormones gibberellin （GA） and abscisic acid （ABA） play key roles in the regulation of plant fertility （Kasembe, 1967; Stein, 1971; Jacobsen and Olszewski, 1991; Zhu et al., 2010）, but the detailed mecha- nisms largely remain unknown so far.

Expression of NAC1 up-stream regulatory region and its relationship to the lateral root initiation induced by gibberellins and auxins

A 1050 bp up-stream regulatory fragment of the transcription factor gene NAC1in Arabidopsis thaliana was isolated using polymerase chain reaction (PCR) based techniques. Thefragment was used to substitute the 35S promoter of the pBI121 plasmid to construct abate-glucuronidase gene (GUS) expression system. The construct was introduced into tobacco(Nico-tiana tabaccum) plants by the Agrobacterium-med\aled transferring method. GUS expressionpattern was studied by using the transgenic lines. The results showed that the GUS driven by theNAC1 up-stream regulatory region was specifically expressed in the root meristem region, basal areasof the lateral root primordium and the lateral roots. The GUS expression was induced by3-indolebutyric acid (IBA)and gibberellins (GA_3 and GA_(4+7)). The results indicated that theup-stream regulatory fragment of NAC1 responded to plant hormones. The fragment might be involved inboth auxins and gibberellins signaling in promoting the development of lateral roots.

Histological,transcriptomic,and gene functional analyses reveal the regulatory events underlying gibberellin-induced parthenocarpy in tomato

Gibberellin(GA)is one of the major plant hormones that promote parthenocarpy,a highly valuable agronomic trait.Here,we demonstrated that exogenous GA3application triggered the formation of parthenocarpic fruits with smaller size but unchanged shape in tomato(Solanum lycopersicum).These fruits exhibited a thicker pericarp,undeveloped ovules,and few jelly tissues,leading to smaller locules with empty cavities.Histological investigation showed that GA treatment produced more cell layers with larger cells in the pericarp,suggesting its promotion in both cell division and expansion.Transcriptomic analyses between GA-3and mock-treated unpollinated ovaries/fruits identified a large number of differentially expressed genes related to hormones,cell division,cell expansion,and transcription factors,implying that they coordinately regulated parthenocarpy conferred by GA.In particular,the downregulation of five reported repressors of tomato parthenocarpy,including two auxin signaling components,AUXIN RESPONSE FACTOR5(SlARF5)and Sl ARF7,and three MADS-box genes,TOMATO APETALA3(TAP3),TOMATO PISTILLATA(TPI),and AGAMOUS-LIKE6(SlAGL6),after GA treatment might play a key role in this process.Furthermore,we found that the knockdown of a GA signaling factor SlMYB33,which was depressed by GA treatment,induced parthenocarpic fruit set in tomato,an effect that might have been achieved by enhancing GA biosynthesis and decreasing the expression of some repressors of tomato parthenocarpy.Thus,our results provide a basis for understanding the regulatory mechanism of GA in tomato parthenocarpy.

OsbZIP01 Affects Plant Growth and Development by Regulating OsSD1 in Rice

As the‘Green Revolution’gene,SD1(encoding GA20ox2),has been widely applied to improve yield in rice breeding.However,research on its transcriptional regulation is limited.Here,we identified a transcription factor OsbZIP01,which can suppress the expression of SD1 and regulate gibberellin(GA)biosynthesis in rice.Knockout mutants of OsbZIP01 exhibited increased plant height,while the overexpression lines showed a semi-dwarf phenotype and diminished germination rate.Furthermore,the semi-dwarf phenotype of OE-bZIP01,was caused by the reduced internode length,which was accompanied by a thin stem width.The predominant expression of OsbZIP01 was observed in leaves and sheaths.OsbZIP01 protein was localized in the nucleus and showed transcriptional repression activity.In addition,OsbZIP01 could directly bind to the promoter of the OsSD1 gene,and inhibit its transcription.The semi-dwarf phenotype of OE-bZIP01 could be rescued by exogenous GA_(3).Meanwhile,the bzip01 sd1 double mutant showed a shorter shoot length compared with the wild type,indicating that OsbZIP01 regulated plant growth mainly through the GA biosynthesis pathway.Collectively,OsbZIP01 negatively regulates GA biosynthesis by restraining SD1 transcription,thereby affecting plant growth and development.

Dc GA20ox2 and DcGA2ox1 alter endogenous gibberellin contents and adjust lignin accumulation in carrot

Gibberellins(GAs)are a class of plant hormones that can affect plant growth and development.GA-oxidases are rate-limiting enzymes,which play a direct role in GA accumulation in plants.However,the roles of GA-oxidase on carrot(Daucus carota L.)taproot development are still unclear.In this study,two GA-oxidase genes,DcGA20ox2 and DcGA2ox1,were identified in carrot.Transgenic carrot plants were obtained by using Agrobacterium-mediated genetic transformation method.The results showed that overexpression of DcGA20ox2 significantly promoted the accumulation of active GAs in carrot,increased plant height,generated more branches,and enhanced xylem development.Overexpression of DcGA2ox1 significantly reduced the total contents of active GAs compared with the control group,resulting in a dwarf phenotype and markedly increased lignin content of the transgenic carrot.The expression profiling showed that the genes of GA metabolic pathway responded to the negative feedback regulation mechanism.At the same time,the expression of most genes in lignin biosynthesis and polymerization process was up-regulated,corresponding to the massive accumulation of lignin.These findings indicated that DcGA20ox2 and DcGA2ox1affected carrot growth and development by regulating the levels of endogenous GAs.The results from current work might shed light on further studies aimed to regulate lignification in carrot and other crops.

EjGASA6 promotes flowering and root elongation by enhancing gibberellin biosynthesis

The Gibberellic Acid-stimulated Arabidopsis(GASA)gene family is involved in the regulation of gene expression and plant growth,development,and stress responses.To investigate the function of loquat GASA genes in the growth and developmental regulation of plants,a loquat EjGASA6 gene homologous to Arabidopsis AtGASA6 was cloned.EjGASA6 expression was induced by gibberellin,and ectopic transgenic plants containing this gene exhibited earlier bloom and longer primary roots since these phenotypic characteristics are related to higher gibberellin content.Transcriptome analysis and qRT-PCR results showed that the expression levels of GA3ox1 and GA20ox1,which encode key enzymes in gibberellin biosynthesis,were significantly increased.Furthermore,we confirmed that EjGASA6 could promote the expression of GA20ox1 via the luciferase reporter system.Overall,our results suggest that EjGASA6 promotes blooming and main-root elongation by positively regulating gibberellin biosynthesis.These findings broaden our understanding of the role of GASAs in plant development and growth,and lay the groundwork for future research into the functions of EjGASA6 in regulating loquat growth and development.

Overexpression of PbrGA2ox1 enhances pear drought tolerance through the regulation of GA_(3)-inhibited reactive oxygen species detoxification and abscisic acid signaling

Drought stress is a devastating natural disaster driven by the continuing intensification of global warming,which seriously threatens the productivity and quality of several horticultural crops,including pear.Gibberellins(GAs)play crucial roles in plant growth,development,and responses to drought stress.Previous studies have shown significant reductions of GA levels in plants under drought stress;however,our understanding of the intrinsic regulation mechanisms of GA-mediated drought stress in pear remains very limited.Here,we show that drought stress can impair the accumulation of bioactive GAs(BGAs),and subsequently identified PbrGA2ox1 as a chloroplast-localized GA deactivation gene.This gene was significantly induced by drought stress and abscisic acid(ABA)treatment,but was suppressed by GA_(3)treatment.PbrGA2ox1-overexpressing transgenic tobacco plants(Nicotiana benthamiana)exhibited enhanced tolerance to dehydration and drought stresses,whereas knock-down of PbrGA2ox1 in pear(Pyrus betulaefolia)by virus-induced gene silencing led to elevated drought sensitivity.Transgenic plants were hypersensitive to ABA,and had a lower BGAs content,enhanced reactive oxygen species(ROS)scavenging ability,and augmented ABA accumulation and signaling under drought stress compared to wild-type plants.However,the opposite effects were observed with PbrGA2ox1 silencing in pear.Moreover,exogenous GA_(3)treatment aggravated the ROS toxic effect and restrained ABA synthesis and signaling,resulting in the compromised drought tolerance of pear.In summary,our results shed light on the mechanism by which BGAs are eliminated in pear leaves under drought stress,providing further insights into the mechanism regulating the effects of GA on the drought tolerance of plants.

Identification of P-type plasma membrane H^(+)-ATPases in common wheat and characterization of TaHA7 associated with seed dormancy and germination

The P-type plasma membrane(PM)H^(+)-ATPases(HAs)are crucial for plant development,growth,and defense.The HAs have been thoroughly characterized in many different plants.However,despite their importance,the functions of HAs in germination and seed dormancy(SD)have not been validated in wheat.Here,we identified 28 TaHA genes(TaHA1-28)in common wheat,which were divided into five subfamilies.An examination of gene expression in strong-and weak-SD wheat varieties led to the discovery of six candidate genes(TaHA7/-12/-14/-16/-18/-20).Based on a single nucleotide polymorphism(SNP)mutation(C/T)in the TaHA7 coding region,a CAPS marker(HA7)was developed and validated in 168 wheat varieties and 171 Chinese mini-core collections that exhibit diverse germination and SD phenotypes.We further verified the roles of the two allelic variations of TaHA7 in germination and SD using wheat mutants mutagenized with ethyl methane sulphonate(EMS)in‘Jimai 22’and‘Jing 411’backgrounds,and in transgenic Arabidopsis lines.TaHA7 appears to regulate germination and SD by mediating gibberellic acid(GA)and abscisic acid(ABA)signaling,metabolism,and biosynthesis.The results presented here will enable future research regarding the TaHAs in wheat.

Effect Evalution of Xiulijing on Fruit Rust Control of Golden Delicious Apple

[Objectives]The paper was to evaluate the effectiveness of Xiulijing in the management of rust in Golden Delicious apple cultivation.[Methods]The Golden Delicious apple was utilized as the test material.During the young fruit stage,various dilutions of Xiulijing were applied,specifically at concentrations of 200,400,600,800,1000,and 2000 times.Additionally,both the application of a 3000-fold dilution of GA 4+7 and the practice of fruit bagging were designated as the control treatments,while a water spray was utilized as the blank control.The fruit rust index and the incidence of rust-infected fruits were evaluated to assess the effectiveness of control measures for fruit rust in Golden Delicious apple cultivation.[Results]The application of Xiulijing,with dilutions ranging from 200 to 2000 times,exhibited a significant impact on reducing both the fruit rust index and the incidence of rust-infected fruits in Golden Delicious apples.The application of a 1000-fold dilution of Xiulijing resulted in a statistically significant reduction in both the fruit rust index and the incidence of rust-infected fruits,with reduction rates of 21.1%and 31.5%,respectively.Among the various strategies for the prevention and control of apple rust,fruit bagging emerged as the most effective method.The other two technical measures demonstrated significant control effects;however,no substantial differences were observed between them.[Conclusions]The biological control agent Xiulijing is more deserving of promotion and application in practical production due to its significantly superior economic benefits,safety,and stability.

RNAi Plasmid Construction of PttGA 20-oxidase Gene

[Objective] Genetic Engineering technology was used to regulate the expression of PttGA20-oxidase gene thus restrained plant height growth and internode elongation for cultivating dwarfed plant.[Method] Based on the RNAi principle,the gene specific sequences of PttGA20-oxidase in the antisense and sense orientations interrupted by a gene sequence from GUS were cloned into a binary vector pBI121.The selection marker gene npt Ⅱ was replaced with bar gene to RNAi plasmid.[Result] After undergone different endonuclease restrictions,the constructed constraint vector released different segments whose sizes were similar to that of target segment,which demonstrated that the RNAi plasmid of PttGA20-oxidase gene was successfully constructed.[Conclusion] The experiment provided a new way for culturing dwarfed plant.

Experiments on decolourization with 122-resin for extraction of gib-berellins from solid medium under different conditions

The extracting liquid of gibberellins (Gibberella fujikuroi) from solid medium for was decolorized separately with 75%, 95% alcohol, and distilled water in static adsorption and vibrating way for different durations. The results showed that the content of GA3 in efflux extracted with alcohol is 10% higher than that with distilled water. With the increase of the durations of extraction, the content of GA3 increases and the dissolution of pigments also increase. For extraction of GA3 cultured in solid medium, the best decolorizing result was obtained when it was extracted with 75%-95% alcohol in static way for 1-3 h, or in a vibrating way for 30-60 min, and then to decolor in a static way for 30-60 min in 122 resin column.

Effects of Nitric Oxide on the Germination of Wheat Seeds and Its Reactive Oxygen Species Metabolisms Under Osmotic Stress

Effects of sodium nitroprusside (SNP), a nitric oxide (NO) donor, on the germination and metabolism of reactive oxygen species were surveyed in wheat (Triticum aestivum L.) seeds. Germination of wheat seeds and even the elongation of radicle and plumule were dramatically promoted by SNP treatments during the germination under osmotic stress. Meanwhile, activities of amylase and EP were enhanced, thus leading to the degradation of storage reserve in seeds. After osmotic stress was removed, higher viability of wheat seeds was also maintained. In addition, the activities of CAT, APX and the content of proline were increased by SNP treatment simultaneously, but activities of LOX were inhibited, and both of which were beneficial for improving the antioxidant capacity during the germination of wheat seeds under osmotic stress. It was also shown that the increase of the activity of amylase induced by SNP in embryoless half-seeds of wheat in the beginning period of germination (6 h) might be indirectly related to GA(3).

From model to alfalfa:Gene editing to obtain semidwarf and prostrate growth habits

Alfalfa(Medicago sativa L.)is a nutritious forage crop with wide ecological adaptability.The molecular breeding of alfalfa is restricted by its heterozygous tetraploid genome and the difficult genetic manipulation process.Under time and resource constraints,we applied a more convenient approach.We investigated two MtGA3ox genes,MtGA3ox1 and MtGA3ox2,of Medicago truncatula,a diploid legume model species,finding that MtGA3ox1 plays a major role in GA-regulated plant architecture.Mutation of neither gene affected nitrogenase activity.These results suggest that MtGA3ox1 can be used in semidwarf and prostrate alfalfa breeding.Based on the M.truncatula MtGA3ox1 sequence,MsGA3ox1 was cloned from alfalfa,and two knockout targets were designed.An efficient CRISPR/Cas9-based genome editing protocol was used to generate msga3ox1 mutants in alfalfa.We obtained three lines that carried mutations in all four alleles in the T0 generation.Fifteen clonal plants were vegetatively propagated from each transgenic line using shoot cuttings.The plant height and internode length of msga3ox1 null mutants were significantly decreased.The number of total lateral branches,leaf/stem ratio and crude protein content of aerial plant parts of msga3ox1 mutants were significantly increased.Thus,we obtained semi-dwarf and prostrate alfalfa by gene editing.